Purification of Guggul by Ayurvedic Process (Shodhana)
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Indian Journal of Traditional Knowledge Vol. 17 (2), April 2018, pp. 391-395 Purification of Guggul by Ayurvedic process ( Shodhana ), estimation of Guggulsterone E & Z before and after purification by HPLC analysis Prince Kumar Pal 1, Goli Penchala Prasad 1, Gajji Babu 1, Dev Nath Singh Gautam 2 & Narendra Kumar Singh 3* 1Regional Ayurveda Research Institute for Skin Disorders (Under CCRAS, Ministry of Ayush, Govt. of India), Vijaywada-520 015, Andhra Pradesh, India; 2Department of Rasa Shastra, Faculty of Ayurveda, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221 005, Uttar Pradesh, India; 3Pharmacy Ayurveda Research Laboratory, Rajiv Gandhi South Campus, Banaras Hindu University, Barkachha, Mirzapur-231 001, Uttar Pradesh, India. E-mail: [email protected] Received 21 February 2017, revised 24 July 2017 Guggul is one of the Ayurvedic product in great demand, used for over thousands of years and high status for its versatile use in several ailments. The two most important pharmacological properties of guggul are its anti-inflammatory and hypolipidaemic actions. The present study was undertaken to find out the effect of three different media of purification (water, cow’s urine and Triphala Kashay a (decoction of three myrobalans)) of raw guggul on its markers Guggulsterone E & Z at different heating temperature during Shodhana (purification) process. The guggul purified by different methods was found to be quite variable in reference to physico-chemical parameters and its marker compounds Guggulsterone E & Z by HPLC analysis at similar chromatographic conditions. The Guggulsterone E & Z was found in the order: water > cow’s urine > Triphla Kashaya (0.29 w/w, 0.24 w/w and 0.16 w/w, respectively at 85 °C to 95 °C). But the same raw guggul purified with same media like water, cow’s urine and Triphala Kashay a at temperature 60 °C to 70 °C showed the presence of high quantity of Guggulsterone E & Z (0.49 w/w, 0.45 w/w and 0.30 w/w for water, cow’s urine and Triphala Kashay a, respectively), as compared to the quantity of Guggulsterone E & Z found at temperature 85 °C to 95 °C. Keywords: Guggul , Shodhana , Guggulsterone E & Z, HPLC. IPC Int. Cl. 8: A61K 36/00, A61K 9/00, A61K 45/06, B01D 53/34, A61K 39/395, C07D 457/14 Guggul , an oleo-gum-resin is obtained from agent 2,6,9-11 . Two triterpenes, myrrhanol A and Commiphora wightii (Arnott.) Bhandari (syn. myrrhanone A of guggul have been shown to possess Commiphora mukul ) of family Burseraceae. It is potent anti-inflammatory activity 12 . In addition to its mentioned in classic Ayurvedic texts to be highly hypolipidaemic and anti-inflammatory activities, efficacious in the treatment of various disorders guggul has been reported to possess antioxidant 13-15 , particularly obesity, lipid disorders, rheumatism, antibacterial, antifungal 16,17 , antitumor 18,19 , thyroid arthritis, cardiovascular and neurological diseases1. The stimulatory 20,21 and cardioprotective activities 22,23 . hypolipidaemic, anti-atherosclerotic 2-4 and anti- Guggulsterone is also known to inhibit tumor cell inflammatory activities of guggul have been proliferation and induced apoptosis 24 . The beneficial investigated in detail and gugulipid (an ethylacetate effect of guggul in coronary heart disease has been extract of guggul ) was commercially marketed in India associated with increased fibrinolysis and reduced in 1988 as a hypolipidaemic drug 5. The effectiveness of platelet aggregation 25,26 . guggul for treating arthritis has been well Though, guggul used for the treatment of several demonstrated 6-9. It has been effectively used to reduce diseases, yet it is mentioned in Ayurvedic texts that the possible cause of inflammation by removing administration of raw guggul may sometimes lead to deposits of waste and toxic material from the body skin rashes, irregular menstruation, diarrhoea, including mineral deposition from the joints, thereby headache, mild nausea and very high doses cause liver acting as anti-rheumatic and anti-inflammatory toxicity 27 . In order to overcome the several unwanted _________ effects of raw guggul , a number of traditional *Corresponding author purification processes ( Shodhana ) have been described 392 INDIAN J TRADIT KNOWLE, VOL. 17, NO. 2, APRIL 2018 in Ayurveda with different ‘ dravyas ’, i.e ., fluids. These thoroughly. The mixture was allowed to stand for purification processes not only remove the adverse next 12 h. The content was then heated using gas effects of guggul but also enhance the therapeutic stove and gentle boiling was maintained until drug- activity. It is also mentioned in Ayurvedic texts, that water mixture got reduced to one-forth of its guggul must be purified before being incorporated into original volume. Afterwards boiling mixture was herbal formulations 28 . There are large numbers of cooled at room temperature and solid residue was commercial polyherbal anti-inflammatory formulations allowed to settle down. The mixture was strained available in the market in which guggul is a chief through muslin cloth and the solid residue was ingredient 29 . However, no study has been done to rinsed with boiling potable water. The strained explore the possible changes during the purification mixture was once again filtered and filtrate was process and its probable effect on level of key pooled to get ‘ Triphla Kashaya ’. constituents, Guggulsterone E & Z. Therefore, the The foreign matter like sand and stone we re present study was carried out to purify raw guggul by removed from guggul . The guggul (1 Kg) was broken using three different Shodhana Dravyas (purifying into small pieces and taken in the pottali (bundled in substances), viz. water, cow’s urine and Triphla cotton cloth). It was hung in Dola Yantra Kashaya as mentioned in Ayurvedic texts and to (an instrument used to impregnate medicine in evaluate the possible changes in the level of key liquids) in Triphla Kashaya (2 L) taken in an iron constituents Guggulsterone E & Z during its vessel. Triphla Kashaya was heated at a temperature purification process. between 85 °C- 95 °C until all the guggul passed into the fluid through the cotton cloth. The residue in the Materials and methods pottali was discarded and fluid was filtered and boiled Chemicals and solvents until it was converted in form of semi-solid mass . The Guggulsterone E & Z were purchased from Sigma- mass was dried in tray dryer at 50 °C and pounded Aldrich Co. LLc., India. All other solvents, chemicals with a pestle in a stone mortar 29 . The same procedure and reagents used were of analytical grade. Distilled was followed for the purification of guggul with water wherever used was filtered from Millipore filter. cow’s urine and water at different temperature for comparative analytical determination. Procurement of raw material Crude drug guggul gum was procured from Determination of physico -chemical parameters Radhika Enterprises, New Delhi, India and Loss on drying, total ash, acid insoluble ash, water authenticated on the basis of macroscopic, soluble extractive value and resin content were microscopic characters and physico-chemical determined before and after the purification of guggul parameters as described in the Ayurvedic as per the Ayurvedic Pharmacopoeia of India 30 . Pharmacopoeia of India. Other crude drugs such as Haritaki (Terminalia chebula Retz.), Vibhitaki Quantitative estimation of Guggulsterone E & Z (Terminalia bellirica Gaertn Roxb.) and Amalaki Test solution was prepared by refluxing of 3.0 g of (Emblica officinalis Gaertn. syn: Phyllanthus emblica L.) sample with 50 mL of acetonitrile on water bath for (these three drugs are constituents of Triphla in equal 30 min. It was cooled and filtered through Whatman proportions) were procured from the local market of filter paper No. 45 in a volumetric flask and volume Vijayawada, Andhra Pradesh, India and authenticated was made up to 100 mL by adding sufficient amount as per the parameters described in the Ayurvedic of acetonitrile. Small amount of sample was filtered through millipore filter (0.45 µm pore size) in a vial Pharmacopoeia of India. Cow’s fresh urine was 31 collected from Goshala of Vijayawada Andhra and placed in sample holder of HPLC instrument . Pradesh, India. HPLC instrument (Agilent Technologies 1260 infinity) was equipped with stainless steel column of Shodhana of guggul dimension 25 cm x 4.6 mm and packed with C-18 Authenticated crude drugs Haritaki , Vibhitaki and silica gel (5 µm). A filtered and degassed mixture of Amalaki were coarsely powdered and mixed in equal 45 volume of acetonitrile and 55 volume of water was quantities. This mixed powder called Triphla’ (500 g) used as mobile phase with flow rate 2 mL/min. was transferred to an extraction vessel. Potable water Samples (20 µL) were injected in the column and (8.0 L) was added to the extraction vessel and mixed temperature was maintained at 25 °C. UV-visible PAL et al. : SHODHANA OF GUGGUL BY AYURVEDIC PROCESS 393 Table 1 — Physico-chemical evaluation of raw guggul and purified guggul at temp. 85 °C to 95 °C Parameters Raw guggul Purified guggul Purified guggul with Purified guggul with with water cow’s urine Triphala Kashaya Loss on drying 2.30 2.11 2.04 1.57 Total ash 14.32 10.34 9.67 9.88 Acid insoluble ash 9.23 5.32 4.99 5.03 Water soluble extract 45.31 35.10 33.34 39.02 Resin content 36.45 24.91 24.09 18.86 Table 2 — Physico-chemical evaluation of raw guggul and purified guggul at temp. 60 °C to 70 °C. Parameters Raw guggul Purified guggul with Purified guggul with cow’s Purified guggul with Triphala water urine Kashaya Loss on drying 2.30 2.21 2.01 1.89 Total ash 14.32 11.04 8.97 8.68 Acid insoluble ash 9.23 4.92 5.19 6.03 Water soluble extractive 45.31 37.15 34.34 38.02 value Resin content 36.45 29.91 26.09 20.97 spectrophotometer was used as detector and sample compared to the quantity of Guggulsterone E & Z found was detected at 242 nm 31 .